Compositions comprising a retinoid and an NFkB-inhibitor and their methods of use

ABSTRACT

A composition including a retinoid, an NFκB-inhibitor, and a cosmetically-acceptable topical carrier is provided. Methods of treating the skin are also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.12/911,038 filed Oct. 25, 2010, the disclosure of which is herebyincorporated by reference in its entirety.

FIELD OF THE INVENTION

A composition comprising a retinoid, an NFκB-inhibitor and acosmetically acceptable topical carrier is provided. The composition isuseful for topical application to the skin.

BACKGROUND OF THE INVENTION

Retinoids are known to treat different skin conditions such as acne orphoto-aging. However retinoids used for topical application, such asretinoic acid, retinaldehyde or retinol can, in certain instances resultin redness, itching, stinging, skin scaling or other manifestations ofirritation. The inventors have considered that a possible solution todecrease irritation is to use lower concentrations of retinoids in thetopical composition. However compositions with reduced levels ofretinoids can, understandably, have reduced efficacy.

The inventors have now surprisingly discovered that a particular classof anti-inflammatory compounds, agents that inhibit the celltranscription factor, nuclear kappa-B (NFκB) can be combined withretinoids in a manner to provide a surprisingly large and unexpectedsynergistic boost in retinol activity, as measured by the expression ofthe CRAPBII gene.

SUMMARY OF THE INVENTION

In one aspect, the invention provides a composition comprising aretinoid, an NFκB-inhibitor, and a cosmetically-acceptable topicalcarrier, wherein the amount of retinoid in the composition is no morethan about 0.075% by weight of the composition.

In another aspect, the invention provides a method of treating skin thatincludes topically applying the above composition to mammalian skin.

Other features and advantages of the present invention will be apparentfrom the detailed description of the invention and from the claims.

DETAILED DESCRIPTION OF THE INVENTION

It is believed that one skilled in the art can, based upon thedescription herein, utilize the present invention to its fullest extent.The following specific embodiments are to be construed as merelyillustrative, and not limitative of the remainder of the disclosure inany way whatsoever.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention belongs. Unless otherwise indicated, anamount, percentage, or concentration refers to an amount, percentage, orconcentration by weight (i.e., % (W/W). Unless stated otherwise, allranges are inclusive of the endpoints, e.g., “from 4 to 9” includes theendpoints 4 and 9.

As used herein, “topically applying” means directly laying on orspreading on outer skin, the scalp, or hair, e.g., by use of the handsor an applicator such as a wipe, roller, or spray.

As used herein, “cosmetically acceptable” means that the ingredients theterm describes are suitable for use in contact with tissues (e.g., theskin or hair) without undue toxicity, incompatibility, instability,irritation, allergic response, or the like.

As used herein, the term “treating” or “treatment” means the treatment(e.g., alleviation or elimination of symptoms and/or cure) and/orprevention or inhibition of the condition (e.g., a skin condition).

Compositions of the present invention are suitable for treatingmammalian skin, for example for improving various signs of skin aging,such as firmness, texture, or the appearance of wrinkles. As usedherein, “skin in need of improving the signs of aging” means a skin thatis, but not limited to, sagging, loose, lax, rough, wrinkly, thinned, oruneven. Improving the signs of aging means improving the firmness of theskin, improving the texture of the skin, improving the appearance oflines or wrinkles in skin, improving the skin tone, or the treatment ofexternal aggressions in skin.

As used herein, “improving the firmness of skin” means the enhancing ofthe firmness or elasticity of the skin, preventing the loss of firmnessor elasticity of skin, or preventing or treating sagging, lax and looseskin. The firmness or elasticity of the skin can be measured by use of acutometer. See Handbook of Non-Invasive Methods and the Skin, eds. J.Serup, G. Jemec & G. Grove, Chapter 66.1 (2006). The loss of skinelasticity or firmness may be a result of a number of factors, includingbut not limited to aging, disease, hormonal changes, trauma,environmental damage, or the result of an application of cosmetics tothe skin.

As used herein, “improving the texture of skin” means the smoothing ofthe surface of the skin to remove either bumps or crevasses on the skinsurface.

As used herein, “improving the appearance of lines or wrinkles in skin”means preventing, retarding, arresting, or reversing the process ofwrinkle and fine line formation in skin.

As used herein, “treatment of external aggressions in skin” means thereduction or prevention of the damage from external aggressions in skin.Examples of external aggressions include, but are not limited to, damageto the skin from the use of cleansers (e.g., topical cleanserscontaining surfactants), make-up, shaving as well as environmentaldamage such as from UV light (e.g., sun damage from sunlight or damagefrom non-natural sources such as UV lamps and solar simulators), ozone,exhaust, pollution, chlorine and chlorine containing compounds, andcigarette smoke. Effects of external aggressions on the skin include,but are not limited to, oxidative and/or nitrosative damage to andmodifications on lipids, carbohydrates, peptides, proteins, nucleicacids, and vitamins. Effects of external aggressions on the skin alsoinclude, but are not limited to, loss of cell viability, loss oralteration of cell functions, and changes in gene and/or proteinexpression.

As used herein, the term “lightening the skin” refers generally tolightening, brightening, whitening, and/or evening of the skin tone,skin color, and/or shade of skin, and/or to the reduction in sallowness,and/or to the lightening and/or fading of hyperpigmented marks and/orlesions including, but not limited to, pigmented spots, melanin spots,age spots, sun spots, senile lentigos, freckles, lentigos simplex,pigmented solar keratosis, seborrhoeic keratosis, melasma, acne marks,post-inflammatory hyperpigmentation, lentigines, ephelides, combinationsof two or more thereof and the like. In certain embodiments, “lighteningthe skin” also refers to increased skin radiance, glow, translucencyand/or luminescence and/or obtaining a more radiant, glowing,translucent or luminous skin tone appearance or a less yellow or sallowskin tone. In certain preferred embodiments, “lightening the skin”refers to lightening and evening the skin tone, increasing skin radianceand/or lightening age spots.

As used herein, the term “skin in need of skin lightening treatment”refers generally to skin that exhibits one or more property selectedfrom the group consisting of: skin having a measured Individual TypologyAngle (ITA) value below 41 as determined per the COLIPA GUIDELINE:GUIDELINE FOR THE COLORIMETRIC DETERMINATION OF SKIN COLOUR TYPING ANDPREDICTION OF THE MINIMAL ERYTHEMAL DOSE (MED) WITHOUT UV EXPOSUREpublished in 2007, which is incorporated herein by reference and furtherdescribed below, darkened and/or sallow skin, including skin darkened byUV, skin with uneven skin tone, or skin with one or more hyperpigmentedmarks and/or lesions including, but not limited to, pigmented spots,melanin spots, age spots, sun spots, senile lentigos, freckles, lentigossimplex, pigmented solar keratosis, seborrhoeic keratosis, melasma, acnemarks, post-inflammatory hyperpigmentation, lentigines, ephelides,combinations of two or more thereof and the like. In the COLIPAguidelines, skin color is defined function of the ITA value as: verylight skin>55; Light skin 41-55, Intermediate 28-41, and Tan skin<28. Incertain preferred embodiments, “skin in need of skin lightening” refersto individuals with a skin having an ITA value of less than 41, such asabout 40 or less, about 35 or less, about 30 or less, or more preferablyabout 28 or less. In certain other preferred embodiments, the presentinvention is directed to compositions and methods for use on skin inneed of skin lightening treatment selected from sallow and/or darkenedskin. In certain other preferred embodiments, the present invention isdirected to compositions and methods for use on skin in need of skinlightening treatment selected from the group consisting of age spots,freckles, marks left after acne, and combinations of two or morethereof.

Compositions of the present invention are also suitable for treatingacne. As used herein, “treating acne” refers to a mitigating, reducing,preventing, improving, or eliminating the presence or signs of disordersresulting from the actions of hormones and other substances on thesebaceous glands and hair follicles, typically leading to clogged poresand the formation of inflammatory or non-inflammatory lesions on theskin. Specifically, it relates to the treatment or prevention ofblemishes, lesions, or pimples, pre-emergent pimples, blackheads, and/orwhiteheads. As used herein, a “pre-emergent pimple” is an inflamedfollicle that are not visually apparent on the surface of the skin withthe naked eye (e.g., as a lesion).

As used herein, “cosmetic” refers to a beautifying substance orpreparation which preserves, restores, bestows, simulates, or enhancesthe appearance of bodily beauty or appears to enhance the beauty oryouthfulness, specifically as it relates to the appearance of tissue orskin.

As used herein, “cosmetically effective amount” means an amount of aphysiologically active compound or composition sufficient for treatingone or more signs of skin aging or acne, but low enough to avoid seriousside effects. The cosmetically effective amount of the compound orcomposition will vary with the particular condition being treated, theage and physical condition of the end user, the severity of thecondition being treated/prevented, the duration of the treatment, thenature of other treatments, the specific compound or product/compositionemployed, the particular cosmetically-acceptable carrier utilized, andlike factors.

Retinoid

Compositions of the present invention include at least one retinoid. Asused herein, “retinoid” means a compound from a class of compoundsstructurally similar to Vitamin A, such as those characterized by thestructure below:

wherein R represents a functional group such as CH₂OH (retinol), CHO(retinal), CO₂H (retinoic acid), CH₂OCOCH (retinyl acetate). Otherderivatives are suitable as well, e.g. other esters such as retinylpalmitate, amine derivatives, and the like. In one embodiment, theretinoid is selected from retinol, retinal, retinoic acid, retinylacetate, and retinyl palmitate. In a preferred embodiment, the retinoidis retinol.

The inventors have found that retinoid levels can be maintained atrelatively low levels in the composition, yet provide a surprisinglylarger than expected efficacy. In certain embodiments of the invention,the amount of retinoid (e.g., retinol) in the composition is no morethan about 0.075% by weight of the composition (e.g., no more than about0.0026 moles per liter of composition or no more than about 247,500International Units of Vitamin A per 100 grams of composition). Incertain embodiments, the amount of retinoid in the composition is fromabout 0.01% to about 0.075% by weight of the composition (e.g., fromabout 0.000349 to about 0.0026 moles per liter of composition), such asfrom about 0.01% to about 0.06% by weight of the composition (e.g., fromabout 0.000349 to about 0.00209 moles per liter of composition). In apreferred embodiment, the composition comprises retinol in the aboveconcentration ranges.

NFκB-Inhibitor

Compositions of the present invention include at least oneNFκB-inhibitor. As used herein, “NFκB-inhibitor” means a compound thatinhibits the cell transcription factor nuclear kappa-B (NFκB). In oneembodiment, the NFκB-inhibitor, when tested according to theNFκB-INHIBITION TEST as defined below, has a Percent NFκB Inhibition ofat least about 35%, preferably at least about 55%, more preferably atleast about 70%, most preferably at least about 90%, when tested at aconcentration that is preferably from 1 microgram per milliliter toabout 100 micrograms per milliliter. That is, the compound demonstratesthe recited Percent NFκB Inhibition at at least one concentration in therange of 1 microgram per milliliter to 100 micrograms per milliliter.The compound need not provide the recited Percent NFκB Inhibition at allconcentrations from 1 microgram per milliliter to 100 micrograms permilliliter, but provides the recited Percent NFκB Inhibition at at leastone concentration in this range.

In a preferred embodiment, the NFκB-inhibitor has a Percent NFκBInhibition of at least about 35%, preferably at least about 55%, morepreferably at least about 70%, most preferably at least about 90%, whentested at a concentration of 10 micrograms per milliliter.

The NFκB-INHIBITION TEST is conducted in the following manner. FB293cells, a stable transfected human epithelial cell line containing thegene reporter for NF-κB are used. They may be obtained from, e.g.,Panomics (Fremont, Calif.). FB293 are plated at a density of 5×10⁴cells/mL in a suitable medium, e.g., Dulbecco's modified Eagle's medium(DMEM) supplemented with 10% fetal bovine serum (Invitrogen, San Diego,Calif.). The FB293 cells are stimulated with 100 ng/mL of Tumor NecrosisFactor-α (TNFα, available from Sigma-Aldrich of St Louis, Mo.) in thepresence of the test sample. Separately, a control sample is testedwherein no test sample is applied. Following a 24-hour incubation at 37°C. with 5% CO₂, cells are lysed with 40 μl of reporter lysis buffer(Promega, Madison, Wis.). A 20-μl aliquot of the lysate is assayed usinga luciferase assay kit (Promega) and counted for 10 s in a Lmaxluminometer (Molecular Devices, Sunnyvale, Calif.) with the datarepresented as the relative light unit/second. Percent NFκB Inhibitionof the test sample is calculated as:NFκB Inhibition=[1−(L _(sample) /L _(control))]*100where L_(sample) is the luminescence of the sample and L_(control) isthe luminescence of the control.

In one embodiment, the NFκB-inhibitor is selected from the groupconsisting of the following compounds: substituted resorcinols,(E)-3-(4-methylphenylsulfonyl)-2-propenenitrile (such as “Bay 11-7082,”commercially available from Sigma-Aldrich of St. Louis, Mo.),tetrahydrocurcuminoids (such as Tetrahydrocurcuminoid CG, available fromSabinsa Corporation of Piscataway, N.J.), extracts of Paulowniatomentosa wood and combinations thereof. In another embodiment, theNFκB-inhibitor is selected from the group consisting of substitutedresorcinols, tetrahydrocurcuminoids, and combinations thereof.

In one embodiment, the NFκB-inhibitor comprises an extract of Paulowniatomentosa wood. Paulownia is a genus of plants native to Asia which hasspread gradually to Europe and the USA. In Japan, Paulownia is calledkiri which refers specifically to one species, Paulownia tomentosa, alsocalled “Princess Tree.” Other names which are commonly used are “empresstree”, “Foxglove Tree”, “Royal Paulownia”, “Pao tong” (in China) and“Odong-Namoo” (in Korea). The scientific name is “Paulownia tomentosa”with a number of synonyms reported in various literature, i.e.“Paulownia imperialis”, “Paulownia recurva”, and “Bignonia tomentosa”.Paulownia tomentosa belongs to the family “Paulowniaceae” or sometimesrefer to “Scrophulariaceae”. The United States Department of Agriculture(plants.USDA.gov) Plant database identifies Princess tree by a uniquesymbol “PATO2”, with Paulownia tomentosa and Paulownia imperialis assynonym names.

Any suitable extracts of Paulownia tomentosa wood may be used. Ingeneral, the wood of the Paulownia tomentosa tree includes wood from thestem, branches, or a combination of both. Suitable extracts of Paulowniatomentosa wood may be derived from wood chips, wood dusts and/or smallcuttings, and the like.

Suitable extracts of Paulownia tomentosa wood may be obtained usingconventional methods including, but not limited to, direct extraction ofmaterial from the wood by grinding, macerating, pressing, squeezing,mashing, centrifuging, and/or processes such as cold percolation,agitation/distillation, microwave assisted extraction, sonication,supercritical/subcritical CO₂ compressed gas extraction with or withoutpolar modifiers, pressurized solvent extraction, accelerated solventextraction, pressurized or normal hot water extraction, surfactantassisted pressurized hot water extraction, oil extraction, membraneextraction, Soxhlet extraction, the gold finger distillation/extractionand/or processes disclosed, for example, in U.S. Pat. Nos. 7,442,391,7,473,435, and 7,537,791 to Integrated Botanical Technologies, LLC,incorporated herein by reference, and the like, or by other methods suchas solvent extraction, and the like. Any of a variety of solventsincluding polar solvents, non-polar solvents, or combinations of two ormore thereof may be used in methods of comprising solvent extraction.Suitable polar solvents include polar inorganic solvents such as waterand the like, polar organic solvents such as alcohols and correspondingorganic acids, for example C₁-C₈ alcohols including methanol, ethanol,propanol, butanol, and the like and organic acids, including aceticacid, formic acid, propanoic acid, and the like, polyols and glycols,including C₁-C₈ polyols/glycols and the like, and combinations of two ormore thereof. Suitable non-polar solvents include non-polar organicsolvents such as alkanes, including C₁-C₈ alkanes, cycloalkanes,including C₁-C₈ alkanes, alkyl ethers, including C₁-C₈ alkyl ethers,Petroleum ethers, ketones, including C₁-C₈ ketones, methylene chloride,ethyl acetate, xylene, toluene, chloroform, vegetable oil, mineral oiland the like. In another embodiment extraction may be obtained bynon-polar solvents described above or supercritical fluid extractionwith or without a polar modifier such as C₁-C₈ alcohols, water, C₁-C₈polyols/glycols or C₁-C₈ organic acids. In certain preferredembodiments, the extract of the invention is a polar extract prepared bypulverizing the wood and extracting using a polar solvent having adielectric constant value of between 1 and 100 at 20° C., preferably adielectric constant of a value between 4 and 60 at 20° C., morepreferably a dielectric constant of a value between 4 and 50 at 20° C.,and even more preferably a dielectric constant of a value between 4 and40 at 20° C. Examples of preferred polar solvents include C₁-C₈alcohols, C₁-C₈ polyols/glycols, C₁-C₈ organic acids, water andcombinations of two or more thereof having a dielectric constant valueof between 1 and 100, preferably between 4 and 60, and more preferablybetween 5 and 40 at 20° C., including, but not limited to, thosesolvents and combinations of solvents having the desired dielectricconstant value as disclosed in “Dielectric Constants of Some OrganicSolvent-Water Mixtures at Various Temperatures,” Akerlof, Gosta; JACS,Vol. 54, No. 11 (November 1932), pp. 4125-4139, incorporated herein byreference. In certain preferred embodiments, the polar extract isextracted using one or more C₁-C₈ alcohols, C₁-C₈ polyols, C₁-C₈glycols, and combinations of two or more thereof. In certain morepreferred embodiments, the extract is extracted using one or more C₁-C₄alcohols, C₁-C₄ polyols, and/or C₁-C₄ glycols. In certain more preferredembodiments, the extract is prepared using a solvent comprisingmethanol, ethanol, or a combination thereof with or without presence ofwater. In more preferred embodiment, the extract is prepared usinganhydrous alcohol or reagent grade denatured alcohol and dried Kiri wooddust agitating at room temperature for 3 days. In certain preferredembodiments, the extract may be further refined by charcoal (alsoreferred to as active carbon) treatment.

In certain embodiments, the Paulownia tomentosa extract may be preparedto be essentially free of certain materials. In one embodiment, theextract is essentially free of Ursolic acid, beta-Sitosterol, or both.

In certain embodiments, the composition may additionally includeextracts from other parts of Paulownia tomentosa, for example, one ormore of the bark, leaves, roots, fruits, seeds, or flowers. In otherembodiments, the composition is essentially free from extracts of othernon-wood parts of Paulownia tomentosa.

In a preferred embodiment, the NFκB-inhibitor is a substitutedresorcinol. Resorcinol is a dihydroxy phenol compound (i.e., 1,3dihydroxybenzene) having by the following structure:

As used herein, “substituted resorcinol” means resorcinol comprising atleast one substituent in the 2, 4, 5, or 6 position. Thus, thesubstituted resorcinol may have as few as one and as many as foursubstituents. Positions 1 and 3 of the substituted resorcinol comprise—OH groups, as shown above.

It is highly preferred that all of the substituents of the substitutedresourcinol are free of phenyl (—C6H5 aromatic) moieties. In certainembodiments, all of the substituents are free of aromatic moieties (withor without heteroatoms).

In another embodiment, it is preferred that all of the substituents ofthe substituted resorcinol are free of ketone functionalities (carbonylsbonded to two other carbon atoms).

In certain preferred embodiments, all of the substituents of thesubstituted resorcinol are free of both phenyl functionalities andketone functionalities.

In certain preferred embodiments, the substituted resorcinol comprisesat least one substituent comprising 5 to 11 carbon atoms, preferably 5to 10 carbon atoms, more preferably 5 to 9 carbon atoms, most preferably5 to 8 carbon atoms. In certain other embodiments, at least onesubstituent comprises an alkyl group, such as one having the number ofcarbon atoms described above. The alkyl group is preferably unsaturated.

In certain embodiments, the 4 position of the resorcinol is substituted,and, in certain embodiments, only the 4 position is substituted. Inanother embodiment, the 4 position is substituted with an akyl group. Incertain preferred embodiments, the substituted resorcinol comprises asingle substituent at the 4 position that comprises an alkyl group. Incertain other preferred embodiments, the substituted resorcinolcomprises a single substituent at the 4 position that consists of analkyl group directly bonded to the benzene ring.

Particularly suitable substituted resorcinols include 4-hexyl resorcinoland 4-octylresorcinol, particularly 4-hexyl resorcinol. The structuresof 4-hexylresorcinol and 4-octylresorcinol are shown below:

4-Hexyl resorcinol is commercially available as “SYNOVEA HR” fromSytheon of Lincoln Park, N.J. 4-Octylresorcinol is commerciallyavailable from City Chemical LLC of West Haven, Conn.

In certain embodiments, the substituted resorcinol comprises at leasttwo substituents in the 2, 4, 5, or 6 positions. Such substituents mayoptionally be linked to form a ring, such as a cyclic aliphatichydrocarbon optionally comprising heteroatoms such as sulfur or oxygen.Such a linked substituent may comprise 5 to 10 carbon atoms, e.g., 8 to10 carbon atoms, and optionally include 1 to 3 heteroatoms. Examples ofsuitable substituted resorcinols comprising cyclic aliphaticsubstituents joining the 2 and 3 positions include Zearalanone andβ-Zearalanol:

Zearalanone and β-Zearalanol are commercially available from SigmaChemicals of St. Louis, Mo.

In certain other embodiments, the substituted resorcinol compriseshalide-containing and/or nitroso-containing substituents. Suitableexamples contain —Cl or —N═O bonded directly to the benzene ring. Thesesubstituents may exist for example in the 2 and 4, 2 and 6, or 4 and 6positions. An example of a dihalide-substituted resorcinol is2,6-dichlororesorcinol. An example of a dinitroso-substituted resorcinolis 2,4-dinitrososorcinol:

2,6-Dichlororesorcinol and 2,4-Dinitrososorcinol are available from CityChemical LLC of West Haven, Conn.

Substituted resorcinols are prepared by means known in the art, forexample, using techniques described in U.S. Pat. No. 4,337,370, thecontents of which are incorporated herein by reference.

The substituted resorcinols may have any suitable molecular weight. Incertain embodiments, the molecular weight of the substituted resorcinolranges between about 175 and about 300.

CRAPBII Expression

In one embodiment of the invention, the concentration of NFκB-inhibitorin the composition is selected to provide, in combination with theretinoid, at least about a 15% synergy in CRAPBII expression as measuredby the CRAPBII EXPRESSION TEST, described below. Such a combinationadvantageously allows for the use of relatively low levels of retinoids,which are sometimes irritating, while maintaining a high degree ofretinoid activity. In humans, the CPABPII gene encodes the synthesis ofcellular retinoic acid-binding protein. As such, expression of theCRABPII gene is directly related to and strongly correlated withretinoid efficacy. The inventors have found that the presence ofNFκB-inhibitor in a composition containing a retinoid improves orpotentiates the beneficial effects of retinoid as measured by CRAPB IIexpression.

The degree of synergy shown in CRAPBII expression, “% Synergy,” for aparticular combination of retinoid and NFκB-inhibitor can be determinedusing the CRAPBII EXPRESSSION TEST. The CRAPBII EXPRESSSION TEST isconducted in the following manner. One centimeter-diameter ex vivo skinexplants are prepared from skin abdominal biopsy after plastic surgery.The skin explants are maintained in KGM Gold™ culture mediumsupplemented with amphotericinB, 0.125 μg/ml at 37° C., in a watersaturated atmosphere for the 48 hour duration of the test. The explantsare placed in a conventional test plate, epidermal surface oriented up,and in sufficient culture medium to nearly but not completely immersethe sample (i.e., the epidermal surface protrudes from the upper surfaceof the medium). Test composition is applied to the epidermal surfaceprotruding from the culture medium. After 48 hours, using conventionaltechniques known to those skilled in the art, the explants are removed,from which epidermal mRNA is extracted and expression of CPABPII gene ismeasured by Quantitative real time PCR (QRT-PCR) using a suitablesequence of oligonucleotides.

A water-in-oil emulsion base containing the desired retinoid is used forthe test composition. The NFκB-Inhibitor may optionally be included inthe test composition or mixed separately into the culture medium. Forinstance, depending on the relative penetration rate of theNFκB-Inhibitor, it may be desirable to include it in the testcomposition or the culture medium to eliminate penetration rate as avariable in the test. If contained in the culture medium, theNFκB-Inhibitor need not penetrate into the epidermis from the protrudingepidermal surface, but rather may penetrate the epidermis from theunderlying culture medium. If the NFκB-Inhibitor is included in the testcomposition, the NFκB-Inhibitor must penetrate into the epidermis fromthe protruding epidermal surface.

The concentration of water in the test composition can be adjusted toaccommodate sufficient levels of retinol (and NFκB-Inhibitor, if presentin the test composition). An example of a suitable oil-in water emulsionbase that may be used for the test composition is shown in Table 1below:

TABLE 1 Suitable Oil-In Water Emulsion Base for CRAPBII EXPRESSSION TESTConcentration Tradename INCI (wt. percent) Water, Demineralized Water83.4 Carbopol Ultrez 10 Carbomer 0.4 Edeta BD chelating agent DisodiumEDTA 0.1 Nipagin M Methylparaben 0.2 Propylparaben Propylparaben 0.15Phenoxyethanol Phenoxyethanol 0.5 Sodium Hydroxide Sodium Hydroxide 0.1Simulsol 165 PEG-100 Stearate; Glyceryl 2.0 Stearate Nipanox BHT BHT 0.1Lanette 16 Cetyl Alcohol; Stearyl 1.0 Alcohol; Myristyl Alcohol DUB ININIsononyl Isononanoate 7.0 Propylene Glycol Propylene Glycol 5.0 AscorbicAcid Ascorbic acid 0.05 Crystalline

The CRAPBII expression is determined (i) individually for a given amountof retinoid, (ii) individually for a given amount of NFκB-Inhibitor, and(iii) for a mixture of the retinoid at amount (i) and the NFκB-Inhibitorat amount (ii). The % Synergy is calculated with the formula below:% Synergy=100%×[[CRAPBII _(mixture)/(CRAPBII _(retinoid) +CRAPBII_(NFκB-Inhibitor))]−1]

While the concentration of NFκB-Inhibitor may be selected to provide atleast about 15% synergistic CRABPII expression for the composition ofthe invention, in certain embodiments, the level of NFκB-Inhibitor maybe selected to provide a higher level of synergy, such as at least about50%, or at least about 80% synergy, in CRABPII expression.

The concentration of NFκB-Inhibitor may, in one embodiment, be selected,for example, using the results of the NFκB-INHIBITION TEST. For example,one may estimate a suitable concentration by performing theNFκB-INHIBITION TEST at several (e.g., 4 or more) concentrations, asshown in Example 1 below. The concentration of NFκB-Inhibitor may beselected, for example, so as to provide a Percent NFκB Inhibition of atleast about 35%, preferably at least about 55%, more preferably at leastabout 70%, most preferably at least about 90%, when tested at aconcentration of 10 micrograms per milliliter. This concentration ofNFκB-Inhibitor may be combined in the composition with no more thanabout 0.075% by weight retinoid and % Synergy in CRAPBII expressiondetermined, as shown in Example 2 below.

The inventors have observed that, in certain embodiments, synergy inCRAPBII expression can surprisingly be observed at not only a lowconcentration of retinoid (no more than about 0.075% by weight), butalso low concentrations of NFκB-Inhibitor. This is advantageous, sincethe concentration of retinoid can be maintained at a low level, avoidingpossible negative irritation side effects of the retinoid, whileobtaining a potent boost in efficacy from the NFκB-Inhibitor.

In certain embodiments of the invention, the composition includes nomore than about 0.075% by weight of a retinoid, such as retinol, and nomore than about 0.5% by weight of an NFκB-Inhibitor. In a preferredembodiment, the composition includes no more than about 0.075% of aretinoid and about 0.1% to about 0.5% of an NFκB-Inhibitor.

Topical Compositions

The compositions of the present invention are applied topically to humanskin and/or hair. In addition to the NFκB-inhibitor and retinoid, thecomposition further includes a cosmetically acceptable topical carrierthat may be from about 50% to about 99.99%, by weight, of thecomposition (e.g., from about 80% to about 99.9%, by weight, of thecomposition). In a preferred embodiment of the invention, thecosmetically-acceptable topical carrier is or includes water. Thecosmetically-acceptable topical carrier may include one or moreingredients selected from the group consisting of wetting agents,emollients, oils, humectants, and the like. In one embodiment, thecosmetically-acceptable topical carrier is or includes a substrate suchas a non-woven fabric or film material.

The compositions may be made into a wide variety of product types thatinclude but are not limited to lotions, creams, gels, sticks, sprays,ointments, cleansing liquid washes and solid bars, shampoos, pastes,foams, powders, mousses, shaving creams, wipes, patches, hydrogels,film-forming products, facial masks and skin masks, films and make-upsuch as foundations, and mascaras. These product types may containseveral types of cosmetically acceptable topical carriers including, butnot limited to solutions, suspensions, emulsions such as microemulsionsand nanoemulsions, gels, solids and liposomes.

The compositions useful in the present invention can be formulated assolutions. Solutions typically include an aqueous or organic solvent(e.g., from about 50% to about 99.99% or from about 90% to about 99% ofa cosmetically acceptable aqueous or organic solvent). Examples ofsuitable organic solvents include propylene glycol, polyethylene glycol(200-600), polypropylene glycol (425-2025), glycerol, 1,2,4-butanetriol,sorbitol esters, 1,2,6-hexanetriol, ethanol, and mixtures thereof.

Compositions useful in the subject invention may be formulated as asolution comprising an emollient. Such compositions preferably containfrom about 2% to about 50% of an emollient(s). As used herein,“emollients” refer to materials used for the prevention or relief ofdryness, such as by preventing the transepidermal loss of water from theskin. Examples of emollients include, but are not limited to vegetableoils, mineral oils, fatty esters, and the like.

A lotion can be made from such a solution. Lotions typically containfrom about 1% to about 20% (e.g., from about 5% to about 10%) of anemollient(s) and from about 50% to about 90% (e.g., from about 60% toabout 80%) of water.

Another type of product that may be formulated from a solution is acream. A cream typically contains from about 5% to about 50% (e.g., fromabout 10% to about 20%) of an emollient(s) and from about 45% to about85% (e.g., from about 50% to about 75%) of water.

Although it is preferred that the composition of the present inventionincludes water, the composition may alternatively be anhydrous or anointment that includes no water but organic and/or silicone solvents,oils, lipids and waxes. An ointment may contain a simple base of animalor vegetable oils or semi-solid hydrocarbons. An ointment may containfrom about 2% to about 10% of an emollient(s) plus from about 0.1% toabout 2% of a thickening agent(s).

The composition may be formulated as an emulsion. If the topical carrieris an emulsion, from about 1% to about 10% (e.g., from about 2% to about5%) of the topical carrier contains an emulsifier(s). Emulsifiers may benonionic, anionic or cationic. Examples of suitable emulsifiers includethose typically identified as such in the art of personal care andcosmetic formulations.

Lotions and creams can be formulated as emulsions. Typically suchlotions contain from 0.5% to about 5% of an emulsifier(s). Such creamstypically contain from about 1% to about 20% (e.g., from about 5% toabout 10%) of an emollient(s); from about 20% to about 80% (e.g., from30% to about 70%) of water; and from about 1% to about 10% (e.g., fromabout 2% to about 5%) of an emulsifier(s).

Single emulsion skin care preparations, such as lotions and creams, ofthe oil-in-water type and water-in-oil type are well-known in thecosmetic art and are useful in the subject invention. Multiphaseemulsion compositions, such as the water-in-oil-in-water type or theoil-in-water-in-oil type, are also useful in the subject invention. Ingeneral, such single or multiphase emulsions contain water, emollients,and emulsifiers as essential ingredients.

The compositions of this invention can also be formulated as a gel(e.g., an aqueous, alcohol, alcohol/water, or oil gel using a suitablegelling agent(s)). Suitable gelling agents for aqueous and/or alcoholicgels include, but are not limited to, natural gums, acrylic acid andacrylate polymers and copolymers, and cellulose derivatives (e.g.,hydroxymethyl cellulose and hydroxypropyl cellulose). Suitable gellingagents for oils (such as mineral oil) include, but are not limited to,hydrogenated butylene/ethylene/styrene copolymer and hydrogenatedethylene/propylene/styrene copolymer. Such gels typically containsbetween about 0.1% and 5%, by weight, of such gelling agents.

The compositions of the present invention can also be formulated into asolid formulation (e.g., a wax-based stick, soap bar composition,powder, or a wipe containing powder).

The compositions useful in the subject invention may contain, inaddition to the aforementioned components, a wide variety of additionaloil-soluble materials and/or water-soluble materials conventionally usedin compositions for use on skin and hair, at their art-establishedlevels.

Additional Cosmetically Active Agents

In one embodiment, the composition further contains another cosmeticallyactive agent. As used herein, a “cosmetically active agent” is acompound (e.g., a synthetic compound or a compound isolated from anatural source or a natural extract) that has a cosmetic or therapeuticeffect on the skin or hair, including, but not limiting to, anti-acneagents, shine control agents, anti-inflammatory agents (non-NFκBInhibiting), anti-mycotic agents, anti-parasite agents, externalanalgesics, sunscreens, photoprotectors, antioxidants, keratolyticagents, surfactants, moisturizers, nutrients, vitamins, energyenhancers, anti-perspiration agents, astringents, deodorants, firmingagents, anti-callous agents, and agents for hair and/or skinconditioning.

In one embodiment, the agent is selected from, but not limited to, thegroup consisting of hydroxy acids, benzoyl peroxide, D-panthenol, octylmethoxycinnimate, titanium dioxide, octyl salicylate, homosalate,avobenzone, carotenoids, free radical scavengers, spin traps, amines(e.g., neutrol), ceramides, polyunsaturated fatty acids, essential fattyacids, enzymes, enzyme inhibitors, minerals, hormones such as estrogens,steroids such as hydrocortisone, 2-dimethylaminoethanol, copper saltssuch as copper chloride, peptides containing copper such asCu:Gly-His-Lys, coenzyme Q10, peptides, amino acids such as proline,vitamins, lactobionic acid, acetyl-coenzyme A, niacin, riboflavin,thiamin, ribose, electron transporters such as NADH and FADH2, and otherbotanical extracts such as aloe vera, feverfew, oatmeal and derivativesand mixtures thereof. The cosmetically active agent will typically bepresent in the composition of the invention in an amount of from about0.001% to about 20% by weight of the composition, e.g., about 0.005% toabout 10% such as about 0.01% to about 5%.

Examples of vitamins include, but are not limited to, vitamin Bs such asvitamin B3, vitamin B5, and vitamin B 12, vitamin C, vitamin K, anddifferent forms of vitamin E like alpha, beta, gamma or deltatocopherols or their mixtures, and derivatives thereof.

Examples of hydroxy acids include, but are not limited, to glycolicacid, lactic acid, malic acid, salicylic acid, citric acid, and tartaricacid.

Examples of antioxidants include, but are not limited to, water-solubleantioxidants such as sulfhydryl compounds and their derivatives (e.g.,sodium metabisulfite and N-acetyl-cysteine), lipoic acid anddihydrolipoic acid, resveratrol, lactoferrin, and ascorbic acid andascorbic acid derivatives (e.g., ascorbyl palmitate and ascorbylpolypeptide). Oil-soluble antioxidants suitable for use in thecompositions of this invention include, but are not limited to,butylated hydroxytoluene, tocopherols (e.g., tocopherol acetate),tocotrienols, and ubiquinone. Natural extracts containing antioxidantssuitable for use in the compositions of this invention, include, but notlimited to, extracts containing flavonoids and isoflavonoids and theirderivatives (e.g., genistein and diadzein), extracts containingresveratrol and the like. Examples of such natural extracts includegrape seed, green tea, pine bark, and propolis.

Other Materials

Various other materials may also be present in the composition, as knownin the art. These include humectants, pH adjusters, chelating agents(e.g., EDTA), fragrances, dyes, and preservatives (e.g., parabens) suchas those commonly used in cosmetic formulations

The composition and formulations and products containing suchcompositions of the present invention may be prepared using methodologythat is well known by an artisan of ordinary skill.

Methods of Use

Compositions of the present invention may be topically applied tomammalian skin that is in need of treatment for improvement of one ormore signs of skin aging or acne as described above. In one embodiment,the compositions are applied to skin in need of improvement in firmness,texture, or the appearance of lines and wrinkles. The compositions maybe applied to the skin in need of such treatment according to a suitabletreatment regimen, e.g., from as much as, twice per day to as little asonce every three days or so.

In certain embodiments, compositions of the present invention may alsobe useful for treating other need states associated with skin. Forexample, compositions of the present invention may be useful fortreating post-inflammatory hyperpigmentation, for reducing pore size,for reducing sebum production, and for scar mitigation. In certain otherembodiments, compositions of the present invention may be appliedsimultaneously with or within several hours of a mechanical or physicalexfoliant such as a microdermabrasion treatment, or with a chemicalexfoliant or keratolytic agent such as salicylic acid. In certain otherembodiments, compositions of the present invention are applied to mildwounds or post-surgical sites to facilitate healing.

It is believed that one skilled in the art can, based upon thedescription herein, utilize the present invention to its fullest extent.The following specific embodiments are to be construed as merelyillustrative, and not limitative of the remainder of the disclosure inany way whatsoever. The following non-limiting examples furtherillustrate the invention.

Example 1 NFκB-Inhibition

The NFκB-INHIBITION TEST described above was performed on test samplesof Bay 11-7082 (Sigma-Aldrich, St. Louis, Mo.), TetrahydrocurcuminoidsCG (Sabinsa Corporation, Piscataway, N.J.), as well as variousconcentrations of 4-hexylresorcinol. The results are shown in Table 2,in which NF-κB Gene Reporter Activation (Luminescence, L) is reportedfor the test samples and a control sample. Percent NF-κB Inhibition isalso reported.

TABLE 2 Results of NFκB-INHIBITION TEST NF-kB Gene Reporter ActivationPercent NF-kB (Luminescence, L) Inhibition Untreated  1.2 ± 0.3 — TNFα(100 ng/ml) Stimulated, 108.2 ± 8.5  — “L_(control)” TNFα +4-Hexylresorcinol  9.3 ± 0.9 91.4% (50 μg/ml) TNFα + 4-Hexylresorcinol29.3 ± 9.2 72.9% (10 μg/ml) TNFα + 4-Hexylresorcinol 55.1 ± 1.7 50.9% (5μg/ml) TNFα + 4-Hexylresorcinol 106.1 ± 1.9  1.9% (1 μg/ml) TNFα +Tetrahydrocurcuminoids 37.8 ± 2.6 65.1% CG (10 μg/ml) Bay 11-7082 (25μM) 11.3 ± 5.6 89.5%

Bay 11-7082, Tetrahydrocurcuminoids CG and 4-hexylresorcinol showedstrong NF-κB inhibition.

Example 2 4-Hexylresorcinol and Retinol—Expression of CRAPBII

The CRAPBII EXPRESSSION TEST as described above was performed on aseries of test compositions containing retinol, 4-hexyresorcinol, orcombinations of both of these compounds. For this series of tests, theoil-in water-emulsion base described in Table 1 was used. Retinol and/or4-hexyresorcinol were both included in the water-in-oil emulsion base.Values for CRAPBII expression are reported in Table 3 below. Forcombinations, the % Synergy, calculated as described above, is alsoreported.

TABLE 3 CRAPBII EXPRESSION 4-Hexyl- Retinol resorcinol CRABPII % Example(Wt. %) (Wt. %) Expression Synergy Comp. 1 0.04 0 193 — Comp. 2 0.075 0322 — Comp. 3 0.1 0 498 — Comp. 4 0 0.5 4 — Comp. 5 0 1 27 — Ex. 1 0.040.5 387 96 Ex. 2 0.075 0.5 550 69 Ex. 3 0.04 1 232 5 Ex. 4 0.075 1 41419 Comp. 8 0.1 1 225 −57

As shown in Table 3, CRAPBII expression is positively correlated to andquite sensitive to concentration of retinol over the tested range.4-hexylresorcinol used at low amounts generally improved the CRAPBIIexpression, while higher amounts of 4-hexylresorcinol generally reducedCRAPBII expression. Synergistic response was surprisingly observed whenthe concentration of retinol was no more than about 0.075% by weight ofthe composition. Furthermore, the synergy was greatly magnified when theconcentration of 4-hexylresorcinol was about 0.5% by weight(approximately 50 ng/ml).

Example 3 Bay 11-7082 and Retinol—Expression of CRAPBII

The CRAPBII EXPRESSSION TEST was performed for a series of testcompositions containing retinol, Bay 11-7082, or combinations of both.For this series of tests, the oil-in water-emulsion base shown below inTable 4 was used to make the remainder of the test compositions.

TABLE 4 Oil-In Water Emulsion Base Concentration Ingredient (wt.percent) Aqua 55.7123 Ammonium 0.5 Acryloyldimethyltaurate/VP DisodiumEDTA 0.1 Glycerin 5.0 Butylene Glycol 2.0 PEG-8 5.0 Cetaryl alcohol:Ceteareth-20 3.0 Stearyl Alcohol; Ceteareth-20 3.0 EthylhexylMethoxycinnamate 2.0 Isohexadecane 1.5 PPG-15 Stearyl Ether; BHT 4.5Pentaerythrityl Tetraethylhexanoate 7.0 Butyrospermum Parkii (SheaButter) 1.0 Tocopheryl Acetate 0.25 BHT 0.1 Dimethicone 2.0Cyclohexasiloxane; 2.01 Cyclopentasiloxane Polyacrylamide; C13-14Isoparaffin; 2.0 Laureth-7 Nylon-12 3.0 Ascorbic Acid 0.05 Citric Acid0.25 Sodium Hydroxide 0.0277

For this series of tests, retinol was included in the test composition.However, Bay 11-7082 was not included in water-in-oil emulsion base, butrather was included in the culture medium. CRAPBII expression isreported in Table 5 below. For combinations, the % Synergy is alsoreported.

TABLE 5 CRAPBII EXPRESSION Retinol Bay 11-7082 CRABPII % Example (Wt. %)(μM) Expression Synergy Comp. 9 0.04 0 324 — Comp. 10 0 1 84 — Comp. 110 10 115 — Ex. 5 0.04 1 631 54 Ex. 6 0.04 10 347 −21

As shown in Table 5, CRAPBII expression was found to be synergistic forcombinations of retinol with the NF-κB inhibitor Bay 11-7082. While thecombination of 0.04% retinol with 1 μM of BAY 11-7082 showed synergisticCRAPBII expression, no synergy was observed for the combination of 0.04%retinol with 10 μM of Bay 11-7082.

The data demonstrates that the combination of an NF-κB inhibitor and aretinoid in a composition wherein the retinoid is present in aconcentration of no more than about 0.075% by weight of the compositionproduces a surprising and synergistic increase in retinol activity.

I claim:
 1. A method of treating mammalian skin, comprising applying tosaid skin a composition comprising a retinoid, 4-hexyl resorcinol, and acosmetically-acceptable topical carrier, wherein the amount of retinoidin the composition is no more than 0.075% by weight of the compositionand the amount of 4-hexyl resorcinol is no more than about 0.5% byweight of the composition.
 2. The method of claim 1, wherein said skinis in need of improvement in a sign of skin aging.
 3. The method ofclaim 2, wherein said skin is in need of improvement in firmness,texture, the appearance of lines or wrinkles, or the loss of elasticity.4. The method of claim 1, wherein said skin is in need of improvement ina sign of acne.
 5. The method of claim 4, wherein said skin comprisesblemishes, lesions, or pimples, pre-emergent pimples, blackheads, and/orwhiteheads.